The overall objective of this program is to develop an effective cancer vaccine based on rationally designed T-cell epitope analogs, for initial use in the adjuvant setting for non-small cell lung cancer (NSCLC) and colon cancer. Analog epitopes, enhanced for either human leukocyte antigen (HLA) binding or T-cell receptor (TCR) signaling, have been shown to be more effective at breaking immunologic tolerance than the cognate wild-type epitopes. Although encouraging early-phase clinical data has been obtained using a limited number of HLA-A2-restricted epitope analogs, the immunological and clinical potential for vaccines comprised of multiple epitope analogs restricted by multiple HLA supertypes remains to be investigated. In Phase I of the current proposal, well-established Epimmune technologies and processes will be utilized to complete the identification of 32 novel fixed anchor- and heteroclitic-analog epitopes derived from frequently expressed tumor-associated antigens (TAA) and restricted by the four most commonly expressed HLA supertypes. These analog epitopes will then be used to design and construct a series of optimized minigenes (epigenes). Based on testing of these epigenes in HLA transgenic mice and human in vitro antigen presentation assays, a highly immunogenic epigene will be selected for vaccine development. For the Phase II of this project, we will propose to 1) complete the pre-clinical development program required for IND submission, 2) conduct a Phase I clinical study evaluating safety and immunogenicity of the vaccine, and 3) investigate strategies to enhance immunogenicity that could be incorporated into subsequent clinical testing of the vaccine. The specific aims to be proposed for Phase II of this program will represent a subset of the development activities required to execute this plan. PROPOSED COMMERCIAL APPLICATIONS: Recurrence of disease following surgery, with or without adjuvant chemotherapy, remains a serious medical problem for NSCLC and colon cancer patients. This vaccine is designed to induce T-cell responses directed against multiple TAA, in order to prevent or delay disease recurrence due to metastases.